Isolation and Identification of a Nitrite-oxidizing Bacterium from Hepu County

In order to select the strain that can degrade nitrite, the screening plate with nitrite was used as the sole nitrogen source to screen the strain with ability to degrade nitrite. A strain with nitrite degrading capacity was isolated from the sludge of a shrimp-farming pond in Hepu City, Guangxi Zhuang Autonomous Region. The isolated strain was identified based on colonial morphology, physiological and biochemical characteristics, and 16S rRNA sequence. Results showed that the strain could grow well on the culture medium containing 2.3 g/L nitrite. According to the morphological characteristics, nitrogen source requirements and evolutionary tree of the 16S rRNA sequence, the isolated strain was identified as Acinetobacter radioresistens, which was named HPAR132 strain. This study laid the foundation for further investigation of nitrite-oxidizing bacterium HPAR132.

Achieving and maintaining biological nitrogen removal via nitrite under normal conditions

The principal aim of this paper is to develop an approach to realize stable biological nitrogen removal via nitrite under normal conditions. Validation of the new method was established on laboratory-scale experiments applying the sequencing batch reactor（SBR） activated sludge process to domestic wastewater with low C/N ratio. The addition of sodium chloride（NaCI） to influent was established to achieve nitrite build-up. The high nitrite accumulation, depending on the salinity in influent and the application duration of salt, was obtained in SBRs treating saline wastewater. The maintenance results indicated that the real-time SBRs can maintain stable nitrite accumulation, but conversion from shorter nitrification-denitrification to full nitrification-denitrification was observed after some operation cycles in the other SBR with fixed-time control. The presented method is valuable to offer a solution to realize and to maintain nitrogen removal via nitrite under normal conditions.

Differences in nitrite-oxidizing communities and kinetics in a brackish environment after enrichment at low and high nitrite concentrations

Nitrite accumulation in shrimp ponds can pose serious adverse effects to shrimp production and the environment.This study aims to develop an effective process for the enrichment of ready-to-use nitrite-oxidizing bacteria（NOB）inocula that would be appropriate for nitrite removal in brackish shrimp ponds.To achieve this objective,the effects of nitrite concentrations on NOB communities and nitrite oxidation kinetics in a brackish environment were investigated.Moving-bed biofilm sequencing batch reactors and continuous moving-bed biofilm reactors were used for the enrichment of NOB at various nitrite concentrations,using sediment from brackish shrimp ponds as seed inoculum.The results from NOB population analysis with quantitative polymerase chain reaction（q PCR）show that only Nitrospira were detected in the sediment from the shrimp ponds.After the enrichment,both Nitrospira and Nitrobacter coexisted in the reactors controlling effluent nitrite at 0.1 and 0.5 mg-NO2^--N/L.On the other hand,in the reactors controlling effluent nitrite at 3,20,and 100 mg-NO2^--N/L,Nitrobacter outcompeted Nitrospira in many orders of magnitude.The half saturation coefficients（Ks）for nitrite oxidation of the enrichments at low nitrite concentrations（0.1 and 0.5 mg-NO2^--N/L）were in the range of 0.71–0.98 mg-NO2^--N/L.In contrast,the Ksvalues of NOB enriched at high nitrite concentrations（3,20,and 100 mg-NO2^--N/L）were much higher（8.36–12.20 mg-NO2^--N/L）.The results suggest that the selection of nitrite concentrations for the enrichment of NOB inocula can significantly influence NOB populations and kinetics,which could affect the effectiveness of their applications in brackish shrimp ponds.

Low soil C:N ratio results in accumulation and leaching of nitrite and nitrate in agricultural soils under heavy rainfall

Nitrate (NO-3) and nitrite (NO2-) leaching threatens groundwater quality.Soil C:N ratio,i.e.,the ratio of soil organic carbon to total nitrogen,affects mineralization,nitrification,and denitrification;however,its mechanism for driving soil NO-3and NO-2accumulation and leaching remains unclear.Here,a field investigation in a fluvo-aquic soil and a soil column experiment were performed to explore the relationships between soil C:N ratio and soil NO-3and NO-2leaching in three soil layers (0–20,20–40,and 40–60 cm) under heavy rainfall (rainfall rate>25 mm d-1).The field investigation results showed that both soil NO-3-N and NO-2-N contents decreased exponentially (P<0.001) with increasing soil C:N ratio in each soil layer.Furthermore,negative exponential relationships (P<0.001) were found between soil C:N ratio and both NO-3-N and NO-2-N concentrations in soil solution in each soil layer under heavy rainfall.The soil column divided into three layers was leached with simulated heavy rainfall;the results confirmed negative exponential relationships (P<0.05) between soil C:N ratio and both NO-3-N and NO-2-N concentrations in the leachate from each soil layer.A total of 18 soil samples obtained from three depths at six field sites during the rainy season were used to elucidate the microbial mechanisms induced by soil C:N ratio using high-throughput sequencing and real-time polymerase chain reaction.High abundances of ammonifying bacteria (Flavobacterium,Bacillu,and Pseudomonas),ammonia-oxidizing bacteria (Nitrosospira),and nirS/K gene were observed when soil C:N was low,concomitant with low abundances of NO-2-oxidizing bacteria (Nitrospira) and narG gene.Partial least squares path modeling showed that the high NO-3and NO-2levels at low soil C:N ratio might be attributed to the inhibition of NO-3reduction (i.e.,low narG gene) and NO-2oxidation (i.e.,low Nitrospira) and thus the accumulation of soil NO-3and NO-2,respectively.Therefore,the leaching of NO-2and NO-3in low C:N soils requires more attention during the rainy season.

Immobilization of nitrite oxidizing bacteria using biopolymeric chitosan media

The effects of chitosan characteristics including the degree of deacetylation, molecular weight, particle size, pH pretreatment and immobilization time on the immobilization of nitrite-oxidizing bacteria （NOB） on biopolymeric chitosan were investigated. Nitrite removal efficiency of immobilized NOB depended on the degree of deacetylation, particle size, pH pretreatment on the surface of chitosan and immobilization time. Scanning electron microscope characterization illustrated that the number of NOB cells attached to the surface of chitosan increased with an increment of immobilization time. The optimal condition for NOB immobilization on chitosan was achieved during a 24-hr immobilization period using chitosan with the degree of deacetylation larger than 80% and various particle size ranges between 1-5 mm at pH 6.5. In general, the NOB immobilized on chitosan flakes has a high potential to remove excess nitrite from wastewater and aquaculture systems.

Compositional variations of active autotrophic bacteria in paddy soils with elevated CO_(2) and temperature

Global warming is an increasingly serious ecological problem,we examined how the active autotrophic microbes in paddy soils respond to the elevated CO_(2) and temperature.Here we employed stable isotope probing(SIP)to label the active bacteria using the soil samples from a fully factorial Simulated Climate Change(SCC)field experiment where soils were exposed to ambient CO_(2) and temperature,elevated temperature,elevated CO_(2),and both elevated CO_(2) and temperature.Around 28.9% of active OTUs belonged to ammonia-oxidizing bacteria(AOB)and nitrite-oxidizing bacteria(NOB).Nitrosospira taxa was dominant in all soils and 80.4% of carbon-fixing bacteria under elevated temperature were classified as Nitrosomonas nitrosa.While no labeled NOBs were detected when temperature or CO_(2) were elevated independently,diverse NOBs were detected in the ambient conditions.We found that elevated CO_(2) and temperature had contrasting effects on microbial community composition,while relatively small changes were observed when CO_(2) and temperature were elevated simultaneously.Summarily these results suggest that carbon-fixing bacteria can respond positively to elevated CO_(2) concentrations,but when it’s accompanied with increase in the temperature this positive response could be weakened.Multiple abiotic factors thus need to be considered when predicting how microbial communities will respond to multiple climatic factors.

Evidence for niche differentiation of nitrifying communities in grassland soils after 44 years of different field fertilization scenarios

Long-term nitrogen(N)fertilization imposes strong selection on nitrifying communities in agricultural soil,but how a progressively changing niche affects potentially active nitrifiers in the field remains poorly understood.Using a 44-year grassland fertilization experiment,we investigated community shifts of active nitrifiers by DNA-based stable isotope probing(SIP)of field soils that received no fertilization(CK),high levels of organic cattle manure(HC),and chemical N fertilization(CF).Incubation of DNA-SIP microcosms showed significant nitrification activities in CF and HC soils,whereas no activity occurred in CK soils.The 44 years of inorganic N fertilization selected only 13C-ammonia-oxidizing bacteria(AOB),whereas cattle slurry applications created a niche in which both ammonia-oxidizing archaea(AOA)and AOB could be actively13C-labeled.Phylogenetic analysis indicated that Nitrosospira sp.62-like AOB dominated inorganically fertilized CF soils,while Nitrosospira sp.41-like AOB were abundant in organically fertilized HC soils.The 13C-AOA in HC soils were affiliated with the 29i4 lineage.The 13C-nitrite-oxidizing bacteria(NOB)were dominated by both Nitrospira-and Nitrobacter-like communities in CF soils,and the latter was overwhelmingly abundant in HC soils.The 13C-labeled nitrifying communities in SIP microcosms of CF and HC soils were largely similar to those predominant under field conditions.These results provide direct evidence for a strong selection of distinctly active nitrifiers after 44 years of different fertilization regimes in the field.Our findings imply that niche differentiation of nitrifying communities could be assessed as a net result of microbial adaption over 44 years to inorganic and organic N fertilization in the field,where distinct nitrifiers have been shaped by intensified anthropogenic N input.